温阳化痰散积法对肝癌血管生成及相关蛋白的影响  

The Effect of Warming Yang,Resolving Phlegm,Dispersing Accumulation Mothod on Angiogenesis and Related Proteins in Primary Liver Cancer

作　　者：郭文海[1] 谢和平[1] 邹增城 李永伟[1] GUO Wenhai;XIE Heping;ZOU Zengcheng;LI Yongwei(The Third Affiliated Hospital of Sun Yat-sen University,Guangdong Guangzhou 510630)

机构地区：[1]中山大学附属第三医院,广东广州510630

出　　处：《深圳中西医结合杂志》2024年第8期1-5,共5页Shenzhen Journal of Integrated Traditional Chinese and Western Medicine

基　　金：全国名老中医药专家传承工作室建设项目(140000020132);广东省基础与应用基础研究基金项目(2022A1515011689,2023A1515011937)。

摘　　要：目的:阴阳攻积丸为温阳化痰散积法治疗癥瘕积聚的明朝古方,本研究探讨其化裁后对肝癌细胞株体外肿瘤血管生成和相关蛋白的影响。方法:制备新制攻积丸大鼠含药血清,细胞计数试剂盒(CCK)8检测含药血清对肝癌细胞株HepG2SG的抑制率。蛋白免疫印迹法(Western blot)检测细胞肿瘤血管生成相关蛋白血管内皮钙黏蛋白(VE–cadherin)、波形蛋白(Vimentin)、转化生长因子β2(TGFβ2)、Twist1和CD44的蛋白表达。细胞划痕实验和Transwell法检测HepG2SG细胞迁移侵袭能力。含药血清干预后HepG2SG条件培养基培养人脐静脉内皮细胞(HUVEC),分别检测HUVEC和HepG2SG细胞的体外成管。结果:新制攻积丸含药血清呈浓度依赖性抑制HepG2SG细胞增殖,含药血清组与对照组和空白血清组比较,各浓度抑制率均明显升高(P<0.01);与对照组和空白血清组比较,新制攻积丸含药血清组VE–cadherin,Vimentin、TGFβ2、Twist1和CD44蛋白表达均下调,差异有统计学意义(P<0.01);含药血清组HepG2SG的迁移侵袭能力、HUVEC管状结构生成和HepG2SG的血管生成拟态均较对照组和空白血清组显著减少(P<0.001)。结论:以温阳化痰散积为主的新制攻积丸含药血清可抑制HepG2SG细胞,降低肿瘤迁移侵袭能力,显著下调肿瘤血管生成相关蛋白的表达,可能抑制肿瘤血管生成。Objective Yin Yang Gong Ji Pill is an ancient formula in Ming Dynasty,used to treat the mass and accumulation syndrome with warming Yang,resolving phlegm,and dispersing accumulation.This study explores the effects of the reducing presicription New Gong Ji Pill on tumoRangiogenesis and related proteins in liveRcanceRcell lines in vitro.Methods The drug-containing serum of New Gong Ji Pill was prepared and the inhibition rate of HepG2SG by the drug-containing serum was detected by cell counting kit(CCK)8.Protein expressions of tumoRangiogenesis related proteins such as VE-cadherin,Vimentin,transforming growth factoRβ2(TGFβ2),Twist1 and CD44 were detected by Western blot.Cell scratch assay and Transwell assay were used to detect the migration and invasion ability of HepG2SG cells.Human umbilical vein endothelial cells(HUVEC)cultured with HepG2SG conditional medium and HepG2SG cells were used to detect angiogenesis in vitro,respectively.Results The proliferation of HepG2SG cells was inhibited by the drug-containing serum in a concentration-dependent way.Compared with the control group and the blank serum group,the inhibitory rates of all concentrations in the drug-containing serum group were significantly increased(P<0.01).Compared with the control group and blank serum group,the protein expressions of VE-cadherin,Vimentin,TGFβ2,Twist1 and CD44 in New Gong Ji pill containing serum group were down-regulated,and the differences were statistically significant(P<0.01).The migration and invasion ability of HepG2SG,angiogenesis of HUVEC and angiogenesis mimicry of HepG2SG in drug-containing serum group were significantly reduced compared with the control group and blank serum group(P<0.001).Conclusion The containing serum of New Gong Ji Pill,which mainly contains drugs of expelling phlegm and eliminating accumulation,can inhibit HepG2SG cells,reduce tumoRmigration and invasion ability,significantly down-regulate the expression of tumoRangiogenesis related proteins,and thus inhibit tumoRangiogenesis and angiogenesis mimi

关 键 词：原发性肝癌 阴阳攻积丸 丝甘蛋白聚糖 肿瘤血管生成 

分 类 号：R735.7[医药卫生—肿瘤]